分子杂交法研究肝炎病人血清和肝组织中输血传播病毒

目的 对各型肝炎病人血清和肝组织中输血传播病毒（TTV）核酸检测分析,探讨病毒的致病性。方法 以地高辛为标记物制备TTV DNA探针,斑点杂交法、原位杂交法分别检测血清中及肝组织中TTV DNA。结果 检测103例血清,TTV总阳性率为25．24％（26／103）;甲－戊型肝炎组检出率21．81％（12／55）、非甲－非庚型患者检出率47．37％（9／19）,显著高于正常对照组15％（3／20）。临床可见TTV的单独感染和重叠感染;出现急性、慢性甚至重度肝损伤。12 肝组织可见TTV阳性,阳性颗粒主要见于肝细胞核内。结论 从血清和肝组织证实了TTV的存在,提示这种病毒可能是导致肝脏炎症的一种新病原。     

非甲-非戊型慢性病毒性肝炎患者隐匿性HBV感染的研究

目的 观察非甲-非戊型慢性病毒性肝炎患者隐匿性HBV感染的状况,探讨荧光定量聚合酶链反应(FQ-PCR)技术对隐匿性HBV感染的诊断价值.方法 应用FQ-PCR技术对57例非甲-非戊型慢性病毒性肝炎患者进行了血清、肝组织HBV-DNA定量检测,并将肝组织HBV DNA定量水平与肝脏炎症活动度的关系进行了分析.结果 血清、肝组织HBV DrqA定量阳性分别为13例(22.81%)、22例(38.60%).13例血清HBV DNA定量阳性患者其肝组织定量亦均阳性,但9例肝组织HBV DrqA定量阳性患者其血清定量为阴性,差异有统计学意义(P<0.01);同时13例血清与肝组织定量均阳性患者比较.显示肝组织HBV DNA定量水平显著高于血清定量水平[(6.62±1.21)拷贝,gvs.(4.03±1.06)拷贝/ml,(P<0.01)].肝组织HBV DNA水平与肝脏炎症活动度并无相关性,10例G2,7例G3,5例G4患者HB'q DNA定量分别为(6.13±1.65)拷贝/g、(5.92±1.81)拷贝,g、(5.83±1.89)拷贝/g,(P0.05),但HBV DNA定量阳性患者均为活动性肝脏病变.结论 HBV隐匿性感染是部分非甲-非戊型慢性病毒性肝炎患者的病因.单纯检测血清免疫学标志物对HBV感染诊断存在漏诊,对非甲-非戊型慢性病毒性肝炎患者应用FQ-PCR技术开展血清定量尤其是肝组织中HBV DNA定量检测可提高HBV感染的诊断.对隐匿性HBV感染的慢性病毒性肝炎亦应给予有效的抗病毒治疗.     

原位杂交法检测非甲～庚型肝炎患者肝组织中TT病毒DNA

目的 证实在非甲－庚型病毒性肝炎患者肝组织中TT病毒（transfusion-transmitted virus,TTV)的存在。方法 采用地高辛素标记TTV DNA探针以原位杂交技术对51例血清学病毒标记非甲－戊型、免疫组化检测肝组织中HBsAg、HCV NS3Ag及HGV N55Aag阴性的病毒性肝炎患者石蜡包埋肝组织进行了检测。结果 各型病毒性肝炎肝组织中TTV基因的总检出率为27．5％,其中急性轻型肝炎的检出率为30．8％（4 ／13）,急性重型肝炎为1／8,亚急性重型肝炎为3／7,慢性肝炎为2／6,活动性肝硬化为2／9,慢性重肝肝炎为1／4,原发性肝癌为1／4。TTV DNA表达于肝细胞核或胞质内,以核型多见。在急性肝炎,TTV阳性细胞弥漫分布于肝小叶内,慢性肝炎于汇管区附近较为密集,而在肝硬化病例,阳性细胞在假小叶内多呈片族状不规则分布。结论 在不明原因病毒性肝炎患者血清及肝组织中TTV DNA的检出表明TTV为一种新型的肝炎病毒,TTV为一种嗜肝性病毒。     

检测病毒性肝炎患者血清中SEN病毒及其临床意义

目的：检测病毒性肝炎患者血清中SEN病毒D和H(SENV-D、SENV-H),并探讨其临床意义。方法：采用巢式聚合酶链反应法（nPCR）检测甲、乙、丙、戊型肝炎和非甲－戊型肝炎患者血清中SENV-D和SENV-H DNA。结果：在180例病毒性肝炎患者血清中，SENV-D和SENV-H检出率分别为17.2%(31/180)和5.6%(10/180)，总检出率为18.3%(33/180)、甲、乙、丙、戊型肝炎患者的SENV-D/H检出率高于非甲－戊型肝炎患者。从甲、乙、丙、戊型肝炎和非甲－戊型肝炎患者分离的SENV-D/H核苷酸序列，与SENV-D/H原型株比较，其同源性在94％以上。甲、乙、丙和戊型肝炎患者有无SENV-D/H合并感染，其血清生化学指标无明显差异。结论：SENV-D/H可能不是非甲-戊型肝炎的病原，甲、乙、丙和戊型肝炎患者合并感染SENV-D/H并不加重病情。     

TLMV5′非编码区基因的克隆与序列分析

目的　调查TTV阴性的非甲～庚型肝炎患者中TTV -likeminivirus(TLMV)感染情况 ,对TLMV5′非编码区 (5′NCR)部分基因进行分子克隆与序列分析。方法　采用巢式PCR技术检测5 3例TTV阴性的非甲～庚肝炎患者血清TLMVDNA ,对PCR产物进行克隆、测序和分析。结果　 5 3例病例中TLMVDNA阳性 37例 (6 9 8% ) ,对其中 8株TLMV基因克隆测序 ,并与Takahashi报道的TLMV序列 (GenBankAccessionNo ab 0 2 6 930、ab0 2 6 931)比较 ,其核苷酸序列同源性在 6 4%～ 83%之间。结论　在TTV阴性的非甲～庚肝炎患者中存在TLMV感染。TLMV5′NCR基因变异性较大。TLMV的致病性及其与非甲～庚肝炎的关系尚有待进一步研究     

广州地区肝炎患者血清中庚型肝炎病毒核酸检测和病毒核苷酸测序

目的 了解广州地区肝炎患者中庚型肝炎病毒（HGV）的感染情况及基因型特点。方法 采用逆转录－套式聚合酶链反应（RT－nested PCR)方法检测出肝炎患者血清中HGV RNA,引物位于HGV基因组的非编码区,并对扩增产物进行直接测序。结果 251例急慢性肝炎血清中共检出25例HGV RNA阳性,总阳性率为9．96％,其中56例非甲－戊型肝炎中要出4例阳性（7．14％）,77例乙型肝炎（HB）中检出8例阳性（10．39％）,118例丙型肝炎（HC）中检出13例阳性（11．17％）;2株HGV广州核苷酸之间的同源性为98．0％,与非洲洲的同源性分别为81．7％和84．2％,与美国株的同源性均为91．1％。结论 广州地区肝炎患者中存在HGV感染,2株HGV广州株可能为同一基因型,且与HGV美国株具有较高的同源性。     

TLMV5＇非编码区基因的克隆与序列分析

目的　调查TTV阴性的非甲～庚型肝炎患者中TTV～like mini virus(TLMV)感染情况,对TLMV5＇非编码区(5＇NCR)部分基因进行分子克隆与序列分析。方法　采用巢式PCR技术检测53例T T V阴性的非甲～庚肝炎患者血清TLMV DNA,对PCR产物进行克隆、测序和分析。结果　53例病例中TLMV DNA阳性37例(69.8％),对其中8株TLMV基因克隆测序,并与Takahashi报道的TLMV序列(GenBank Accession No ab 026930、ab026931)比较,其核苷酸序列同源性在64％～83％之间。结论　在T TV阴性的非甲～庚肝炎患者中存在TLMV感染。TLMV5＇NCR基因变异性较大。TLMV的致病性及其与非甲～庚肝炎的关系尚有待进一步研究。     

急性肝炎恢复期血清检测抗戊型肝炎病毒抗体及RNA的临床诊断意义

目的 在急性非甲-戊型肝炎患者中进一步追踪检测不同时期血清抗戊型肝炎病毒(HEV)IgG，以明确临床诊断。方法 用美国Genelabs公司和北京万泰公司抗HEV诊断试剂检测抗HEV，用PCR方法检测HEVRNA，并进行基因序列分析。结果 95例入院首次血清学诊断为急性非甲-戊型肝炎患者中，在住院后11～25d、25～35d分别测定血清抗HEV,16例血清抗HEV阳性(万泰公司)，急性期血清HEVRNA检测10例HEVRNA阳性，经核苷酸序列分析证明，其中4例为Ⅰ型HEV感染，6例为Ⅳ型HEV感染。GenekLbs诊断试剂检测12例抗HEV阳性，7例HEVRNA阳性，其中4例是HEVⅠ型病毒感染，3例是HEV Ⅳ病毒感染。结论 对非甲-戊型肝炎患者进行急性期HEV RNA检测和恢复期抗HEV检测可以进一步明确病原学诊断，在这部分患者中存在HEV不同基因型感染。可能是HEV感染漏诊的原因之一。     

慢性乙型肝炎患者血清和肝组织中HBV DNA含量与庚型肝炎病毒感染的关系

目的：观察慢性乙型肝炎（CH－B）患者血清、肝组织中HBV DNA含量与庚型肝炎病毒（HGV）感染的关系，探讨HGV感染对CH－B患者乙型肝炎病毒（HBV）复制的影响。方法：应用逆转录－聚合酶链反应（RT－PCR）、免疫组织化学法、荧光定量PCR（FQ－PCR）技术方法对56份CH－B患者血清HGV RNA、肝组织HGV Ag、血清及肝组织中HBV DNA含量分别进行了检测，并将血清HGV RNA与肝组织HGV Ag的表达、HGV RNA、HGV Ag阳性与阴性患者HBV DNA含量分别进行了对比研究。结果：血清HGV RNA、肝组织HGV Ag阳性分别为10份（17．9％）、8份（14．3％）。血清HGV RNA阳性与肝组织HGV Ag表达显著相关（P＜0．01），但部分肝组织HGV Ag阴性患者亦有血清HGV RNA表达。血清HGV RNA、肝组织HGV Ag阳性与阴性患者血清及肝组织中HBV DNA含量差异无显著性（P＞0．05）。结论：HGV感染对CH－B患者HBV复制无影响。HGV可在肝脏中复制，但致病性可能较微弱。     

血液透析病人庚型病毒性肝炎感染与丙型病毒性肝炎感染的关系

据报道不同国家血液透析患者庚型病毒性肝炎 (HGV)的发生率为 3%～ 73% ,本文对一组血液透析患者 HGV感染状况及其与丙型病毒性肝炎 (HCV)感染的相关性进行研究。方法　 10 8例血液透析患者 ,分别来自 2个不同的透析单位。男性 6 2例 ,女性 46例 ,平均年龄 5 6 .7岁 ,所有患者除常规实验室检查外 ,均采用巢式 PCR进行 HGV、 HCVRNA检测。结果　 19例 (17.6 % )患者 HGV RNA阳性 ,同样 ,19例 (17.6 % )患者 HCV RNA阳性。其中 8例患者 HGV RNA阳性 (3组 ) ,11例单 HGV RNA阳性 (2组 ) ,11例单纯 HCVNA阳性 (3组 ) ,78例 …     

Retrospective analysis of non-A-E hepatitis: possible role of hepatitis B and C virus infection

In an effort to determine the cause of non-A-E hepatitis, a retrospective study was undertaken on a group of patients with hepatitis but without serological infection markers of hepatitis viruses A-E. A total of 60 patients admitted to Beijing Ditan Hospital during the period of September 1997 and September 1999 were chosen for this study. These patients were diagnosed as either acute or chronic hepatitis, but no serological markers of hepatitis viruses A-E were detected. Since TT virus (TTV), human parvovirus B19 (B19), SEN virus (SENV), and GB virus C/HGV were reported to be associated with hepatitis, attempts were made to detect the presence of these viruses in the sera of patients with non-A-E hepatitis by a nested polymerase chain reaction (nPCR) method. Also, more sensitive nPCR and RT-nPCR methods were used to determine HBV DNA and HCV RNA in these patients. Results derived from these analyses demonstrate that HBV DNA was detected in most of these patients (47/60, 78.3%), suggesting that HBV infection played a major role in occult non-A-E hepatitis and detection of HBV DNA by more sensitive PCR methods such as nPCR should be considered for diagnosis of HBV infection. In addition, HCV RNA was detected in three (5%) of these patients. However, GBV-C (HGV) RNA was not detected, and TTV, B19, and SENV appear not to be associated with non-A-E hepatitis, as the prevalence rates of these viruses in patients with non-A-E hepatitis were similar to those in patients with viral hepatitis A-E. The results from this study indicate that co-infection of TTV or B19 with HBV did not increase the severity of the disease.     

广东地区肝炎患者血清中庚型肝炎病毒RNA的检测

目的了解庚型肝炎病毒（ＨＧＶ）感染在不同临床类型肝炎患者中的存在状况。方法用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）技术对７２１例肝炎患者血清进行了ＨＧＶＲＮＡ检测。结果发现８０例ＨＧＶＲＮＡ阳性，以非甲、乙、丙、丁或戊型肝炎中阳性率为高；慢性丙型肝炎（ＨＣＶ）和慢性乙型混合丙型肝炎（ＨＢＶ＋ＨＣＶ）次之；在慢性重型肝炎和肝硬化时较低。结论庚型肝炎病毒在广东地区现症肝炎患者中有一定程度的流行     

Prevalence of hepatitis B, hepatitis C, GB virus C/hepatitis G and TT viruses in predialysis and hemodialysis patients

Patients with chronic renal failure on hemodialysis have a high risk of infections with viruses such as hepatitis B (HBV), hepatitis C (HCV), GB virus C/hepatitis G (GBV-C/HGV) and TT (TTV) viruses. The prevalence of HBV, HCV, GBV-C/HGV and TTV in patients with chronic renal failure who are on conservative management before entering into a hemodialysis program (predialysis) in comparison with hemodialyzed patients was studied to elucidate whether the high prevalence of these viruses is influenced by that observed in the predialysis stage. The presence of hepatitis B virus surface antigen (HBsAg), HCV RNA, GBV-C/HGV RNA and TTV DNA was analyzed in sera from 80 patients with chronic renal failure (35 on predialysis and 45 on hemodialysis). HBsAg, HCV RNA, GBV-C/HGV RNA and TTV DNA were detected in one (2.8%), six (17.1%), eight (22.5%) and 16 (45.7%) of the 35 patients on predialysis. Two (5.7%) of these patients were coinfected with HCV and GBV-C/HGV, whereas six (17.1%) had GBV-C/HGV and TTV coinfection. In the 45 hemodialyzed patients, HBsAg, HCV RNA, GBV-C/HGV RNA and TTV DNA were detected in one (2.2%), two (4.4%), seven (15.5%) and 26 (57.7%). One (2.2%) patient had HBV and TTV coinfection, two (4.4%) HCV and TTV coinfection whereas four (8.8%) were coinfected with GBV-C/HGV and TTV. No differences regarding age, gender, previous surgery and number of transfusions were found between infected and uninfected patients within and between both groups. In conclusion, the prevalence of the viruses studied in predialysis may influence their prevalence in dialysis units.     

酶联免疫吸附法检测庚型肝炎病毒抗体

目的 探讨抗庚型肝炎病毒（ＨＧＶ）ＩｇＧ与各种病毒感染,丙氨到转氨酶（ＡＬＴ）及ＨＧＶ ＲＮＡ的相关性。方法 用酶联免疫吸附试验（ＥＬＩＳＡ）法检测了３１５例各种肝炎病毒（Ａ￣Ｅ）感染乾和１１７例健康献血员血清的抗－ＨＧＶ ＩｇＧ,并测定其ＡＬＴ,对抗－ＨＧＶ ＩｇＧ阳性的标本用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）法检测ＨＧＶ ＲＮＡ。结果 献血员抗－ＨＧＶ ＩｇＧ的阳性率为３．４２％（４／１１７     

TT病毒与肝炎关系的临床流行病学研究

目的 对闽南地区各种肝炎患者、健康体检者、义力献血员和肝癌患者共４８０例从临床流行病学角度探讨ＴＴ病毒（ＴＴＶ）的致生及其与各种肝炎的关系。方法 采用巢式ＰＣＲＩｇＧ,用ＥＰＩ ＩＮＦＯ６．０软件进行统计分析。结果 ４８０名研究对象中ＴＴＶＤＮＡ的总检出率为２３．９６％。各种胆炎患者的ＴＴＶ总阳性率为２３．９４％,肝癌患者的ＴＴＶ阳性率为２０．６９％,而健康者的ＴＴＶ阳性率为２４．８４％,义务献血     

GB virus C/hepatitis G virus and TT virus infections among high risk renal transplant recipients in India.

BACKGROUND: GB virus C/hepatitis G virus (GBV-C/HGV) and TT virus (TTV) have been widely reported in patients with high parenteral risk such as haemodialysis and renal transplant recipients. The occurrence of these agents in association with hepatitis B virus (HBV) and hepatitis C virus (HCV), in Indian renal transplant recipients, is yet unreported. STUDY DESIGN: Molecular and serological markers of GBV-C/HGV and TTV were examined in addition to those for HBV, HCV and hepatitis D virus (HDV) in a selected group of seventy renal transplant recipients. HGV RNA detection was achieved using primers specific for the 5'NCR and NS5a regions of the genome. Anti-GBV-C/HGV antibody was detected using the mu plate anti-HG env kit (Roche, Germany). TTV DNA PCR was performed using primers specific for the coding region (method A) of the genome. In 50% of patients, TTV DNA was also tested for using primers specific for the non-coding region (method B). Host related factors such as age, alanine aminotransferase (ALT) levels, number of transfusions, haemodialysis sessions, and months following transplantation were also studied. RESULTS: Exposure rates to GBV-C/HGV, TTV (method A), HBV, HCV and HDV were 58.6, 32.9, 52.9, 54.3 and 2.9%, respectively. 'Active' infection as measured by viraemia and/or virus-specific antigenaemia for GBV-C/HGV, TTV, HBV and HCV was 52.9, 32.9, 15.7 and 52.9%, respectively. The majority of GBV-C/HGV and TTV infections were seen as co-infections with other hepatitis viruses. Single infection with GBV-C/HGV and TTV was seen in ten (14.2%) and eight (11.4%) patients, and was not associated with ALT elevation when compared to uninfected blood donors. Using univariate analysis, GBV-C/HGV RNA was significantly associated with > or =20 haemodialysis sessions. TTV DNA occurrence was not associated with any risk factors. CONCLUSIONS: There is a high occurrence of GBV-C/HGV and TTV in this select group of renal transplant recipients in India. These viruses mostly occurred in the context of co-infections with other hepatitis viruses. Long term effects of multiple hepatotropic viral infections need to be carefully documented in such transplant populations.